Abstract: A method of causing the cellular proliferation of one or more cell types contained within a microdroplet having an average volume in the range 4 femtolitres to 10 nanolitres and comprised of an aqueous buffer is provided. It is characterised by the step of incubating the cell(s) inside the droplets in suitable environmental conditions and thereafter detecting the number of cells inside each droplet, characterised in that the microdroplets are suspended in an immiscible carrier fluid further comprising secondary droplets having an average volume less than 25% of the average volume of the microdroplets up to and including a maximum of 4 femtolitres and wherein the volume ratio of carrier fluid to total volume of microdroplets per unit volume of the total is greater than 2:1. The method may be employed for example with microdroplets containing biological cells or with microdroplets containing single nucleoside phosphate such as are prepared in a droplet-based nucleic acid sequencer.

Abstract: Devices, systems, and associated methods are provided for manipulating and/or determining one or more characteristics of cells contained within a biological sample. In particular a device and methods of use thereof are provided, the device comprising a sorting component configured to separate cell-containing microdroplets from empty ones into a population of cell-containing first microdroplets; a microdroplet manipulation component configured to manipulate the first microdroplets using real or virtual electrowetting electrodes, and an optical detection system configured to detect an optical signal from the microdroplets via the one or more detection windows.

Abstract: A method of manipulating microdroplets having an average volume in the range 0.5 femtolitres to 10 nanolitres comprised of at least one biological component and a first aqueous medium having a water activity of aw1 of less than 1 is provided. It is characterised by the step of maintaining the microdroplets in a water-immiscible carrier fluid which further includes secondary droplets having an average volume less than 25% of the average volume of the microdroplets up to and including a maximum of 4 femtolitres and wherein the volume ratio of carrier fluid to total volume of microdroplets per unit volume of the total is greater than 2:1. The method may be employed for example with microdroplets containing biological cells or with microdroplets containing single nucleoside phosphate such as are prepared in a droplet-based nucleic acid sequencer.

Abstract: An apparatus comprising a microfluidic device comprising a microfluidic space configured to contain a plurality of microdroplets; a first light source for illuminating the microfluidic device; a voltage source for supplying a voltage to the microfluidic device in order to generate an electric field across the microfluidic space; a first modulator configured to generate a waveform signal to modulate the electric field applied across the microfluidic space, wherein the first modulator is configured to switch between an active state, in which the electric field is applied across the microfluidic space to hold the plurality of microdroplets; and a non-active state, in which the electric field is not applied across the microfluidic space; and an optical imaging device for generating an image of at least a subset of the microdroplets; wherein the first modulator is further configured to, directly or indirectly, control the first light source such that at least a portion of the light is provided across the microflui

Abstract: A method of detecting the interactions between a biological entity and a molecule, the method comprising providing an array of first microdroplets into a microfluidic chip; wherein each microdroplet contains at least one bead and each bead having a bound photocleavable molecule; providing an array of second microdroplets into the microfluidic chip; wherein each microdroplet contains at least one biological entity; holding the entire first and second arrays of microdroplets; illuminating at least a subset of the first microdroplets containing at least one bead with an illumination source configured to photo-cleave the molecule; subsequently merging at least one subset of the first array of microdroplets with at least one subset of the second microdroplets to form an array of merged microdroplets; and detecting a change in an optical signal from the merged microdroplets using an optical system to indicate the interactions between the biological entity and the molecule.

Abstract: A device for manipulating a microdroplet using optically mediated electrowetting is provided. The device comprising a microfluidic space bounded by: a first composite wall comprising: a first substrate; a first conductor layer on the substrate; a photoactive layer on the first conductor layer; and a first continuous dielectric layer on the photoactive layer having thickness of less than 20 nm; a second composite wall comprising: a second substrate; and a second conductor layer on the substrate.

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.

Abstract: A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5? side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (

Abstract: A device comprising: a first zone comprising an attachment site; a first pathway; a second pathway and a means for creating a second medium comprised of aqueous microdroplets in a carrier; a microdroplet manipulation zone comprising: a first composite wall comprised of a first transparent substrate; a first transparent conductor layer on the substrate; a photoactive layer activated by electromagnetic radiation; and a first dielectric layer on the photoactive layer; a second composite wall comprised of a second substrate; a second conductor layer on the substrate; and optionally a second dielectric layer on the conductor layer; an A/C source; a source of first electromagnetic radiation; means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer; an detection zone disposed downstream of the microdroplet manipulation zone or integral therewith; and a fluorescence or Raman-scattering detection system.

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.

Abstract: A method for selecting cells and/or part(s) of a cell(s) based on at least one characteristic in an EWOD or oEWOD device is provided. The method comprising: i. providing a test panel of microdroplets, comprising at least a medium or a medium and at least one cell; ii. providing at least one reporter panel of microdroplets, containing one or more reporter entities; iii. merging microdroplets of the test panel with microdroplets of the at least one reporter panel to form a panel of merged assay microdroplets; and iv. monitoring the panel of assay microdroplets using a detection system capable of detecting a change in said reporter entity based upon the presence of at least one characteristic, and v.

Abstract: A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5? side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.

Abstract: A device for manipulating many hundreds or thousands of microdroplets into an array using EWOD or oEWOD is provided. The device comprises: i) a chip comprising a first region for receiving and manipulating microdroplets; a second region comprising the array and a plurality of electrowetting pathways leading to the array; ii) a microdroplet source configured to provide microdroplets of a predetermined target diameter; iii) a channel configured to provide fluid communication between the microdroplet source and the first region of the chip; and iv) a pressure source configured to move the microdroplets between the microdroplet source and the first region of the chip. The electrowetting pathways on the chip are centre to centre separated by at least double the predetermined target diameter of the microdroplets from the microdroplet source. Furthermore, the controller is configured to enable synchronous movement of the microdroplets in the electrowetting pathways by application of EWOD or oEWOD force.

Abstract: An apparatus for manipulating one or more microdroplets into an array using EWOD or oEWOD is provided. The apparatus comprises: a) a chip for manipulating microdroplets comprising: i) a plurality of electrowetting pathways leading to the array; and ii) one or more waste electrowetting pathways leading to a waste outlet; b) a detector for detecting one or more microdroplets with a distinct characteristic, said detector configured to obtain a measured dataset relating to the distinct characteristic of the detected microdroplet; c) a storage module configured to store and maintain a stored dataset associated with the characteristic measured by the detector; and d) a controller configured to receive the stored dataset from the storage module and the obtained measured dataset to determine whether the measured dataset is associated with a desired or undesired characteristic.

Abstract: A device is provided which comprises: i) a chip comprising a first region for manipulating a plurality of microdroplets; ii) a microdroplet source for providing the microdroplets; iii) a channel having a distal end, which extends in a first direction into the chip, and a proximal end, which is in fluid communication with the microdroplet source; and iv) a pressure source for moving the microdroplets from the microdroplet source along the channel and into the first region of the chip. The pressure source is configured to enable the movement of the microdroplets from the microdroplet source into the proximal end of the channel at a first velocity. Furthermore, the distal end of the channel is fluted or blunted such that the microdroplets move from the distal end of the channel into the first region of the chip at a velocity which is lower than the first velocity.

Abstract: A method of maintaining at least one component in an aqueous microdroplet dispersed in conditioned oil to form an emulsion is provided.

Abstract: A method of handling an adherent cell in a microdroplet assaying system by conjugating an adherent cell to a microbead is provided. The method 50 comprises the steps of: loading a first plurality of microdroplets into a microfluidic space, wherein each of the first microdroplet 5 contains a microbead 52 and a first fluid; loading a second plurality of microdroplets into the microfluidic space, wherein each of the second microdroplet contains an adherent cell and a second fluid 54; merging the first plurality of microdroplets and the second plurality of microdroplets to form a plurality of merged microdroplets 56, each merged microdroplets containing the first and second fluids, at least one microbead and at least one adherent cell; and10 agitating each of the merged microdroplets 58 to cause the first and second fluids in each of the merged microdroplets to move such that at least one adherent cell adhere to the at least one microbead. [FIG.

Abstract: A method for manipulating a microdroplet of a reaction medium in an immiscible carrier medium with a target molecule bound to a solid support for the purposes of effecting a chemical transformation is provided. It is characterised by the steps of (a) bringing the microdroplet into contact with the solid support under conditions where the microdroplet and solid support are caused to combine, (b) allowing the reaction medium to react with the target molecule and (c) thereafter exerting a force to induce the reaction medium to become detached from the solid support and reform a microdroplet in the carrier fluid. In one embodiment the solid support is a particle, bead or the like.